title: Release of miniantibodies from E. coli cells into the supernatant at low and high cell densities.
abstract: By chemical permeabilization of E. coli cells with detergents and membrane active peptides considerable amounts of the periplasmic anti-PC-miniantibodies can be released. Releases of active miniantibodies of 31%, respectively of 38%, were obtained by exposing low cell density suspensions to nonionic detergents like Triton X-100 and tetraethyleneglycolmonodecylether. At high cell densities releasing levels of 40% and 56% were observed for these detergents. The addition of cationic, membrane active peptides (PMBN, polylysine 115, magainin II and melittin) led to release of up to 20% of active miniantibodies at low cell densities. The excretion of miniantibodies at low cell densities was increased up to 35% by phospholipase A2. Interestingly, the membrane associating properties of the anti-PC-miniantibodies influenced the permeability of the outer membrane and the excretion of beta-lactamase.
authors.affiliation: Institute of Toxicology, ETH, Schwerzenbach, Switzerland.
authors.forename: J L
authors.initials: JL
authors.lastname: Mörbe
authors.forename: D
authors.initials: D
authors.lastname: Riesenberg
mesh_terms: D000907:Antibodies, Bacterial / Q000302:isolation & purification / Q000378:metabolism*; D023181:Antimicrobial Cationic Peptides; D002463:Cell Membrane Permeability / Q000187:drug effects; D003902:Detergents / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development / Q000276:immunology*; D054806:Magainins; D008555:Melitten / Q000494:pharmacology; D009693:Nucleic Acid Hybridization; D010455:Peptides / Q000494:pharmacology; D019897:Periplasm / Q000201:enzymology / Q000502:physiology; D010741:Phospholipases A / Q000494:pharmacology; D054467:Phospholipases A2; D010767:Phosphorylcholine / Q000276:immunology*; D011107:Polylysine / Q000494:pharmacology; D011112:Polymyxin B / Q000031:analogs & derivatives / Q000494:pharmacology; D011506:Proteins / Q000494:pharmacology; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism*; D029867:Xenopus Proteins; D001618:beta-Lactamases / Q000096:biosynthesis
chemical_list: D000907:Antibodies, Bacterial; D023181:Antimicrobial Cationic Peptides; D003902:Detergents; D054806:Magainins; D010455:Peptides; D011506:Proteins; D011994:Recombinant Proteins; D029867:Xenopus Proteins; C055324:phospholipase A2-activating protein; D010767:Phosphorylcholine; C053301:magainin 2 peptide, Xenopus; D008555:Melitten; D011107:Polylysine; C038934:polymyxin B nonapeptide; D010741:Phospholipases A; D054467:Phospholipases A2; D001618:beta-Lactamases; D011112:Polymyxin B